Abstract This study presents a new record of the occurrence of larvae of the genus Ortleppascaris (Sprent, 1978). The nematodes were collected from inside fibrous cysts found in the livers of cane toads, Rhinella marina (Linnaeus, 1758), captured in eastern Brazilian Amazonia. This is the first record of Ortleppascaris larvae in both Brazil and this amphibian host, increasing its distribution in South America, as well as expanding the number of helminths known to infect this toad. The detailed description of Ortleppascaris sp. provides new taxonomic data for these larvae, as well as sequences of the ITS and SSU DNA segments, and the cytochrome oxidase I gene, which will, in time, contribute to a better understanding of the phylogeny of this group of parasites.